3D genome organization coordinates key regulators of lineage specification in mammary epithelial cells [RNASeq_T8]. 3D genome organization coordinates key regulators of lineage specification in mammary epithelial cells [RNASeq_T8]

In order to understand gene regulation of epithelial cells in the mouse mammary gland, we have generated a comprehensive epigenetic atlas from freshly sorted myoepithelial (basal), luminal progenitor (LP) and mature luminal (ML) from adult mice (9 week-old) encompassing gene expression profiling, histone and polymerase II CUT&Tag sequencing, chromatin accessibility via ATAC-seq and chromatin interactions. We further studied the basal compartment by sorting to additional cell type population: Tspan8+ (quiescent basal, QBa) and Tspan8- (active basal, ABa) cells. We then performed gene expression profiling analysis using data obtained from RNA-seq of 2 different cell types Tspan8+ and Tspan8- cells. Overall design: RNA-seq profiles of mouse epithelial mammary gland cell types in the basal compartment: sorted Tspan8+ (quiescent basal, Qba, Lin-/CD29hi/CD24+/Tspan8+) and Tspan8- (active basal, ABa, Lin-/CD29hi/CD24+/Tspan8-) cellular populations from adult mice in triplicate.

为解析小鼠乳腺上皮细胞的基因调控机制，我们从9周龄成年小鼠体内新鲜分选得到肌上皮细胞（myoepithelial）、腔面祖细胞（luminal progenitor, LP）以及成熟腔面细胞（mature luminal, ML），构建了一套全面的表观基因组图谱，涵盖基因表达谱分析、组蛋白与RNA聚合酶II CUT&Tag测序、基于ATAC-seq的染色质开放性检测，以及染色质互作分析。我们进一步对基底细胞群进行细分分选，得到两个亚群：Tspan8+ 静息基底细胞（quiescent basal, QBa）与Tspan8- 活化基底细胞（active basal, ABa）。随后我们基于这两种细胞群体的RNA测序（RNA-seq）数据开展了基因表达谱分析。整体实验设计：对成年小鼠乳腺基底腔室的上皮细胞进行分选，获取Tspan8+（静息基底细胞QBa，Lin-/CD29hi/CD24+/Tspan8+）与Tspan8-（活化基底细胞ABa，Lin-/CD29hi/CD24+/Tspan8-）两个细胞群体，每组设置三次生物学重复，并对其进行RNA测序（RNA-seq）的基因表达谱分析。